Deconvolution Microscopy

Visualization of the MamK protein in magnetotactic bacteria using Deconvolution Microscopy. UC Berkeley 2009.

The MamK protein has been associated with Magnetosome formation and localization. A Magnetosome is a bacterial organelle that contains magnetite crystals that act like the needle of a compass to orient the bacteria in geomagnetic fields. A GFP-MamK fusion protein was created to visualize the localization within the cell (green florescence). To add contrast and a better sense of where MamK was localized, the cell membrane was stained with FM4-64 which causes it to fluoresce red.

To construct the three-dimensional images, bacteria are viewed under a standard florescence microscope and a Z-series of images are taken (or a series of images with the same X and Y coordinates but varying up and down the vertical focus (or Z) axis). After image acquisition, the Z-series of images are then assembled in a software-based process that essentially "re-focuses" an out of focus image. Algorithms are used to extract information out of blurred regions of an image to clean up these regions leaving a clean three-dimensional image with no blurry regions.

The video displays the difference images before and after deconvolution.

To learn more about Magnetotactic Bacteria visit the following link and view recent publications:
http://pmb.berkeley.edu/profile/akomeili#a2

To learn more about Deconvolution Microscopy visit:
http://microscopy.berkeley.edu/courses/TLM/index.html

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